Characterization of the Biological Activities of Recombinant Fusion Protein Green Fluorescent Protein/Human Zona Pellucida Protein 3 (GFP/HZP3)

Despite numerous reports indicating the successful production of bioactive recombinant ZP3, no report has shown the rhZP3 having direct binding activity with human sperm. Recombinant ZP3 generated from our previous study displayed binding activity with human sperm through indirect evidence from hemizona assay (HZA). This present study focused on the production of recombinant ZP3 with direct binding activity with human sperm. Through the application of a pEGFP expression vector, fusion protein GFP/ZP3 was successfully generated and expressed. The expression of GFP/ZP3 was evidenced by RT-PCR and western blot. The fusion protein was partially purified by Ni-NTA affinity column from cell culture medium of stably transfected cells generated from a single cell clone. Immuno-blotting analysis of the fusion protein indicated that the GFP/ZP3 was about 90–92 kD in molecular mass which is close to the sum of the size of GFP and ZP3. Flow cytometry was conducted to evaluate the direct binding activity of fusion protein with human sperm. The GFP/ZP3 showed dose-dependence in the binding assay. GFP tag removal resulted in almost no detectable binding as assayed by flow cytometry, thereby indicating that ZP3 binding was specific. Furthermore, immunofluorescence demonstrated that the fusion protein interacted with human sperm on the acrosome region. In the acrosome reaction assay, fusion protein was proved to be able to induce the sperm acrosome reaction (19.2% ± 3.4% as compared to basal line 10.8% ± 2.3%) at the concentration of 1ug/ml. Therefore, the fusion protein displayed full spectrum biological activity of ZP3, and, most importantly unlike prior studies, this binding activity was elucidated in a direct way

Remedies for Anticipatory Breach of Contract with Two-Sided Asymmetric Information: A Comparison of Legal Regimes

The Law and economics movement has paid a lot of attention to carefully analyzing various doctrines of contract law. Yet, with few exceptions, the doctrine of anticipatory breach seems to have escaped law and economics scholars\u27 scrutiny. Specifically, the question of optimal choice of remedies has escaped scholars\u27 eyes. While traditionally in England the party who files a law suit can get only damages, in the US the party can not only ask for assurances for performance, but also, in appropriate cases, get specific performance. Which regime is better? Can parties opt in and out of those regimes? Is there a legal regime which is superior to both the English and American regimes? In this paper we attempt to start filling in this gap by studying the relationship between various regimes of remedies. Specifically, we start by studying the conditions in which the American legal regime (which grants the non-breaching party an option to choose, in appropriate cases, between specific performance and actual damages) is superior to the English regime (which allows the non-breaching party to seek only actual damages). We then explore a third regime, which as far as we know does not exist, and show that it is unconditionally Pareto Superior to both the English and American legal regimes. Our analysis in this paper informs transactional lawyers of the relevant economic factors they should consider when deciding between remedies in a given anticipatory breach context. We focus on the ex-ante design of the contract in light of new and asymmetric information that the parties anticipate they will gain after they draft the contract. We assume fist, for simplicity, that no renegotiation or investments are involved. We demonstrate the optimal way to design contract clauses which takes advantage of the information that the seller and the buyer receive between the time they enter into the contract and the time of the breach. We present two models. One is for non-market goods and the other is for market-goods. The law is different with respect to the way damages are calculated for these two classes of goods. We thus model both types of transactions. Section two describes the legal background against which we have designed our models. Section three surveys the literature that evaluates contract remedies in the context of anticipatory breach context from an economic perspective. Section four presents two simple models with incomplete two-sided asymmetric information. In section four, we compare the performance of the American legal regime with that of the English one. Section five discusses some interesting extensions meant to approach the first-best allocative efficiency. The appendix provides a more rigorous mathematical demonstration of the model

PABO: Mitigating Congestion via Packet Bounce in Data Center Networks

In today's data center, a diverse mix of throughput-sensitive long flows and delay-sensitive short flows are commonly presented in shallow-buffered switches. Long flows could potentially block the transmission of delay-sensitive short flows, leading to degraded performance. Congestion can also be caused by the synchronization of multiple TCP connections for short flows, as typically seen in the partition/aggregate traffic pattern. While multiple end-to-end transport-layer solutions have been proposed, none of them have tackled the real challenge: reliable transmission in the network. In this paper, we fill this gap by presenting PABO -- a novel link-layer design that can mitigate congestion by temporarily bouncing packets to upstream switches. PABO's design fulfills the following goals: i) providing per-flow based flow control on the link layer, ii) handling transient congestion without the intervention of end devices, and iii) gradually back propagating the congestion signal to the source when the network is not capable to handle the congestion.Experiment results show that PABO can provide prominent advantage of mitigating transient congestions and can achieve significant gain on end-to-end delay